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The stable isotope pyridoxal 5-phosphate-d2 is added to plasma as an internal standard. Meta-phosphoric acid solution is then added to precipitate the proteins. Following sedimentation of the proteins, an aliquot of the clarified supernatant fluid is subjected to separation of pyridoxal 5-phosphate and internal standards from other plasma components by reverse-phase HPLC with quantitation by tandem mass spectrometry.(Unpublished Mayo method)